This application addresses two broad Challenge Areas and three associated specific Challenge Topics including: i) the broad Challenge Area (06) Enabling Technologies, specific Challenge Topic 06-HL- 105: Develop transgenic animal models that are informative for understanding chronic inflammation in humans, and specific Challenge Topic 06-HL-109: Generate reagents for studying lung cell biology and disease progression;and ii) the broad Challenge Area (15) Translational Science, and specific Challenge Topic: 15-DK-102 Develop improved animal models of NIDDK diseases. Cystic Fibrosis (CF) is the most common autosomal recessive disease in Caucasians and is caused by defects in the CFTR chloride channel. Many questions regarding the pathogenesis of this disease remain, in large part due to the lack of a CF animal model that mimics progression of human CF lung disease and exocrine pancreatic insufficiency (CFTR deficient mice lack both these properties). The ferret is an attractive species for modeling CF because: its lung biology is similar to that of humans, its reproductive time is rapid, and it can now be genetically manipulated. The ferret is also the preferred model organism for studying several types of human viral lung infection-most notably infection by influenza virus, which is thought to accelerate the progression of CF lung disease. We have generated a CFTR-deficient model in the ferret by performing somatic cell nuclear transfer with CFTR gene-targeted ferret fibroblasts. Preliminary analysis of CFTR-deficient newborn ferrets demonstrates that they are afflicted by meconium ileus, defects in cAMP-mediated chloride transport in the airway epithelium, and defects in pancreatic excocrine activity-abnormalities also seen in newborn patients with CF. This proposal seeks to characterize pathologies in a newly generated CFTR knockout ferret model, with a focus on enabling its use as a model for studying pathophysiologic mechanisms and therapies in the CF lung. Working closely with Marshall Farms (the major industrial breeder of ferrets in the U.S.), we have established a collaborative relationship that will allow us to achieve these goals in a cost-effective and timely manner. The outcome of this project-initial characterization of CF ferrets, establishment of a new genetic background for easier propagation without intestinal obstruction, and establishment of a corporate partnership for breeding and disseminating this model to the research community-will greatly impact research on the multi-organ aspects of CF disease. Equally important to the characterization of this model is the development of a ferret molecular toolbox. We propose to tackle this issue while also addressing biologic questions about early transcriptional changes within the CF pancreas and lung. 454 GS FLX Titanium deep sequencing technology will be used for transcriptome analyses, giving rise to 3.6-6 Mb of new EST sequence information on the uncharted ferret transcriptome. This information will enable microarray technologies in this species, and the production of antibodies to ferret proteins, thereby greatly facilitating the use of the ferret as a disease model for research. PUBLIC HEALTH RELEVANCE: Cystic Fibrosis (CF) is the most common life-threatening autosomal recessive condition among Caucasians with over $450 million dollars spent on clinical care of CF patients annually in the U.S. alone. Despite the fact that the gene defect responsible for CF was discovered over 20 years ago, a cure for CF has yet to emerge. In large part, this is due to the lack of good animal models that reproduce the human CF disease phenotype on which to both understand the disease process and test therapies. This proposal is focused on the initial characterization and further development of a new animal model of CF in the ferret. This model shows classic signs of human CF disease and may be a valuable resource for the CF research community following completion of the proposed project.